Aerobic glycolysis is a hallmark of metabolic reprogramming that contributes to tumor progression. However, the mechanisms regulating expression of glycolytic genes in neuroblastoma (NB), the most common extracranial solid tumor in childhood, still remain elusive. Crucial transcriptional regulators and their downstream glycolytic genes were identified by integrative analysis of a publicly available expression profiling dataset. In vitro and in vivo assays were undertaken to explore the biological effects and underlying mechanisms of transcriptional regulators in NB cells. Survival analysis was performed by using Kaplan-Meier method and log-rank test. Hepatocyte nuclear factor 4 alpha (HNF4A) and its derived long noncoding RNA (HNF4A-AS1) promoted aerobic glycolysis and NB progression. Gain- and loss-of-function studies indicated that HNF4A and HNF4A-AS1 facilitated the glycolysis process, glucose uptake, lactate production, and ATP levels of NB cells. Mechanistically, transcription factor HNF4A increased the expression of hexokinase 2 (HK2) and solute carrier family 2 member 1 (SLC2A1), while HNF4A-AS1 bound to heterogeneous nuclear ribonucleoprotein U (hnRNPU) to facilitate its interaction with CCCTC-binding factor (CTCF), resulting in transactivation of CTCF and transcriptional alteration of HNF4A and other genes associated with tumor progression. Administration of a small peptide blocking HNF4A-AS1-hnRNPU interaction or lentivirus-mediated short hairpin RNA targeting HNF4A-AS1 significantly suppressed aerobic glycolysis, tumorigenesis, and aggressiveness of NB cells. In clinical NB cases, high expression of HNF4A-AS1, hnRNPU, CTCF, or HNF4A was associated with poor survival of patients. These findings suggest that therapeutic targeting of HNF4A-AS1/hnRNPU/CTCF axis inhibits aerobic glycolysis and NB progression.

中文翻译：

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HNF4A-AS1/hnRNPU/CTCF 轴作为有氧糖酵解和神经母细胞瘤进展的治疗靶点。

有氧糖酵解是代谢重编程的标志，有助于肿瘤进展。然而，神经母细胞瘤（NB）是儿童最常见的颅外实体瘤，调节糖酵解基因表达的机制仍然难以捉摸。通过对公开的表达谱数据集进行综合分析，鉴定了关键的转录调节因子及其下游糖酵解基因。通过体外和体内实验来探索转录调节因子在 NB 细胞中的生物学效应和潜在机制。使用Kaplan-Meier方法和对数秩检验进行生存分析。肝细胞核因子 4 α (HNF4A) 及其衍生的长非编码 RNA (HNF4A-AS1) 促进有氧糖酵解和 NB 进展。功能获得和功能丧失研究表明，HNF4A 和 HNF4A-AS1 促进 NB 细胞的糖酵解过程、葡萄糖摄取、乳酸产生和 ATP 水平。从机制上讲，转录因子 HNF4A 增加己糖激酶 2 (HK2) 和溶质载体家族 2 成员 1 (SLC2A1) 的表达，而 HNF4A-AS1 与异质核核糖核蛋白 U (hnRNPU) 结合，促进其与 CCCTC 结合因子 (CTCF) 的相互作用，导致 CTCF 反式激活以及 HNF4A 和其他与肿瘤进展相关基因的转录改变。阻断HNF4A-AS1-hnRNPU相互作用的小肽或靶向HNF4A-AS1的慢病毒介导的短发夹RNA显着抑制NB细胞的有氧糖酵解、肿瘤发生和侵袭性。在临床 NB 病例中，HNF4A-AS1、hnRNPU、CTCF 或 HNF4A 的高表达与患者较差的生存率相关。这些发现表明，针对 HNF4A-AS1/hnRNPU/CTCF 轴的治疗靶向可抑制有氧糖酵解和 NB 进展。